LOCUS pThermoCas9_ctrl 7467 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses ThermoCas9 and its sgRNA module.
ACCESSION .
VERSION .
KEYWORDS pThermoCas9_ctrl
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7467)
AUTHORS Mougiakos I, Mohanraju P, Bosma EF, Vrouwe V, Finger Bou M,
Naduthodi MIS, Gussak A, Brinkman RBL, van Kranenburg R, van der
Oost J
TITLE Characterizing a thermostable Cas9 for bacterial genome editing and
silencing.
JOURNAL Nat Commun. 2017 Nov 21;8(1):1647. doi: 10.1038/s41467-017-01591-4.
PUBMED 29162801
REFERENCE 2 (bases 1 to 7467)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7467
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(46..62)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(46..62)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(59..81)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 70..86
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(94..124)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 139..160
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(1047..2051)
/codon_start=1
/gene="repB"
/product="RepB replication protein"
/label=repB
/note="from Enterococcus faecalis plasmid pAM-alpha-1"
/db_xref="GI:22652809"
/protein_id="AAN03827.1"
/translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
primer_bind complement(2572..2589)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2743..3331)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2823..2842)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 actataaaac tattgtgcat gggagggaag cttggcgtaa tcatggtcat agctgtttcc
61 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
121 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
181 cgctttccag tcgggaaacc tgtcgtgcca gcccttcaaa cttcccaaag gcgagcccta
241 gtgacattag aaaaccgact gtaaaaagta cagtcggcat tatctcatat tataaaagcc
301 agtcattagg cctatctgac aattcctgaa tagagttcat aaacaatcct gcatgataac
361 catcacaaac agaatgatgt acctgtaaag atagcggtaa atatattgaa ttacctttat
421 taatgaattt tcctgctgta ataatgggta gaaggtaatt actattatta ttgatattta
481 agttaaaccc agtaaatgaa gtccatggaa taatagaaag agaaaaagca ttttcaggta
541 taggtgtttt gggaaacaat ttccccgaac cattatattt ctctacatca gaaaggtata
601 aatcataaaa ctctttgaag tcattcttta caggagtcca aataccagag aatgttttag
661 atacaccatc aaaaattgta taaagtggct ctaacttatc ccaataacct aactctccgt
721 cgctattgta accagttcta aaagctgtat ttgagtttat cacccttgtc actaagaaaa
781 taaatgcagg gtaaaattta tatccttctt gttttatgtt tcggtataaa acactaatat
841 caatttctgt ggttatacta aaagtcgttt gttggttcaa ataatgatta aatatctctt
901 ttctcttcca attgtctaaa tcaattttat taaagttcat ttgatatgcc tcctaaattt
961 ttatctaaag tgaatttagg aggcttactt gtctgctttc ttcattagaa tcaatccttt
1021 tttaaaagtc aatcccgttt gttgaactac tctttaataa aataattttt ccgttcccaa
1081 ttccacattg caataataga aaatccatct tcatcggctt tttcgtcatc atctgtatga
1141 atcaaatcgc cttcttctgt gtcatcaagg tttaattttt tatgtatttc ttttaacaaa
1201 ccaccatagg agattaacct tttacggtgt aaaccttcct ccaaatcaga caaacgtttc
1261 aaattctttt cttcatcatc ggtcataaaa tccgtatcct ttacaggata ttttgcagtt
1321 tcgtcaattg ccgattgtat atccgattta tatttatttt tcggtcgaat catttgaact
1381 tttacatttg gatcatagtc taatttcatt gcctttttcc aaaattgaat ccattgtttt
1441 tgattcacgt agttttctgt attcttaaaa taagttggtt ccacacatac caatacatgc
1501 atgtgctgat tataagaatt atctttatta tttattgtca cttccgttgc acgcataaaa
1561 ccaacaagat ttttattaat ttttttatat tgcatcattc ggcgaaatcc ttgagccata
1621 tctgacaaac tcttatttaa ttcttcgcca tcataaacat ttttaactgt taatgtgaga
1681 aacaaccaac gaactgttgg cttttgttta ataacttcag caacaacctt ttgtgactga
1741 atgccatgtt tcattgctct cctccagttg cacattggac aaagcctgga tttacaaaac
1801 cacactcgat acaactttct ttcgcctgtt tcacgatttt gtttatactc taatatttca
1861 gcacaatctt ttactctttc agccttttta aattcaagaa tatgcagaag ttcaaagtaa
1921 tcaacattag cgattttctt ttctctccat ggtctcactt ttccactttt tgtcttgtcc
1981 actaaaaccc ttgatttttc atctgaataa atgctactat taggacacat aatattaaaa
2041 gaaaccccca tctatttagt tatttgtttg gtcacttata actttaacag atggggtttt
2101 tctgtgcaac caattttaag ggttttcaat actttaaaac acatacatac caacacttca
2161 acgcaccttt cagcaactaa aataaaaatg acgttatttc tatatgtatc aagaatagaa
2221 agaactcgtt tttcgctacg ctcaaaacgc aaaaaaagca ctcattcgag tgctttttct
2281 tatcgctcca aatcatgcga ttttttcctc tttgcttttc tttgctcacg aagttctcga
2341 tcacgctgca aaacatcttg aagcgaaaaa gtattcttct tttcttccga tcgctcatgc
2401 tgacgcacga aaagccctct aggcgcatag gaacaactcc taaatgcatg tgaggggttt
2461 tctcgtccat gtgaacagtc gcatacgcaa tattttgttt cccatactgc attaatgaat
2521 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
2581 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
2641 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
2701 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
2761 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
2821 ataaagatac caggcgtttc cccatggaag ctccctcgtg cgctctcctg ttccgaccct
2881 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
2941 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
3001 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
3061 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
3121 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
3181 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
3241 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
3301 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
3361 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagaa catcctcttt
3421 cttagtttgt atagtcaata aacgaactaa attttcattt ttatcttttt ctagacattt
3481 ttattgtatc gtatttttta ttcaaaaaaa atattgaatt ttaaaaatga tggtgcagta
3541 tgaaggtgta cttagtttat tggttaaaca aagtacgcat caatatattt tttttgatgc
3601 gatttgaatt cgctttcatt atctaaaaca ttcaatcgag gaggtaactt aaatgaagta
3661 taaaatcggt cttgatatcg gcattacgtc tatcggttgg gctgtcatta atttggacat
3721 tcctcgcatc gaagatttag gtgtccgcat ttttgacaga gcggaaaacc cgaaaaccgg
3781 ggagtcacta gctcttccac gtcgcctcgc ccgctccgcc cgacgtcgtc tgcggcgtcg
3841 caaacatcga ctggagcgca ttcgccgcct gttcgtccgc gaaggaattt taacgaagga
3901 agagctgaac aagctgtttg aaaaaaagca cgaaatcgac gtctggcagc ttcgtgttga
3961 agcactggat cgaaaactaa ataacgatga attagcccgc atccttcttc atctggctaa
4021 acggcgtgga tttagatcca accgcaagag tgagcgcacc aacaaagaaa acagtacgat
4081 gctcaaacat attgaagaaa accaatccat tctttcaagt taccgaacgg ttgcagaaat
4141 ggttgtcaag gatccgaaat tttccctgca caagcgtaat aaagaggata attacaccaa
4201 cactgttgcc cgcgacgatc ttgaacggga aatcaaactg attttcgcca aacagcgcga
4261 atatgggaac atcgtttgca cagaagcatt tgaacacgag tatatttcca tttgggcatc
4321 gcaacgccct tttgcttcta aggatgatat cgagaaaaaa gtcggtttct gtacgtttga
4381 gcctaaagaa aaacgcgcgc caaaagcaac atacacattc cagtccttca ccgtctggga
4441 acatattaac aaacttcgtc ttgtctcccc gggaggcatc cgggcactaa ccgatgatga
4501 acgtcgtctt atatacaagc aagcatttca taaaaataaa atcaccttcc atgatgttcg
4561 aacattgctt aacttgcctg acgacacccg ttttaaaggt cttttatatg accgaaacac
4621 cacgctgaag gaaaatgaga aagttcgctt ccttgaactc ggcgcctatc ataaaatacg
4681 gaaagcgatc gacagcgtct atggcaaagg agcagcaaaa tcatttcgtc cgattgattt
4741 tgatacattt ggctacgcat taacgatgtt taaagacgac accgacattc gcagttactt
4801 gcgaaacgaa tacgaacaaa atggaaaacg aatggaaaat ctagcggata aagtctatga
4861 tgaagaattg attgaagaac ttttaaactt atcgttttct aagtttggtc atctatccct
4921 taaagcgctt cgcaacatcc ttccatatat ggaacaaggc gaagtctact caaccgcttg
4981 tgaacgagca ggatatacat ttacagggcc aaagaaaaaa cagaaaacgg tattgctgcc
5041 gaacattccg ccgatcgcca atccggtcgt catgcgcgca ctgacacagg cacgcaaagt
5101 ggtcaatgcc attatcaaaa agtacggctc accggtctcc atccatatcg aactggcccg
5161 ggaactatca caatcctttg atgaacgacg taaaatgcag aaagaacagg aaggaaaccg
5221 aaagaaaaac gaaactgcca ttcgccaact tgttgaatat gggctgacgc tcaatccaac
5281 tgggcttgac attgtgaaat tcaaactatg gagcgaacaa aacggaaaat gtgcctattc
5341 actccaaccg atcgaaatcg agcggttgct cgaaccaggc tatacagaag tcgaccatgt
5401 gattccatac agccgaagct tggacgatag ctataccaat aaagttcttg tgttgacaaa
5461 ggagaaccgt gaaaaaggaa accgcacccc agctgaatat ttaggattag gctcagaacg
5521 ttggcaacag ttcgagacgt ttgtcttgac aaataagcag ttttcgaaaa agaagcggga
5581 tcgactcctt cggcttcatt acgatgaaaa cgaagaaaat gagtttaaaa atcgtaatct
5641 aaatgatacc cgttatatct cacgcttctt ggctaacttt attcgcgaac atctcaaatt
5701 cgccgacagc gatgacaaac aaaaagtata cacggtcaac ggccgtatta ccgcccattt
5761 acgcagccgt tggaatttta acaaaaaccg ggaagaatcg aatttgcatc atgccgtcga
5821 tgctgccatc gtcgcctgca caacgccgag cgatatcgcc cgagtcaccg ccttctatca
5881 acggcgcgaa caaaacaaag aactgtccaa aaagacggat ccgcagtttc cgcagccttg
5941 gccgcacttt gctgatgaac tgcaggcgcg tttatcaaaa aatccaaagg agagtataaa
6001 agctctcaat cttggaaatt atgataacga gaaactcgaa tcgttgcagc cggtttttgt
6061 ctcccgaatg ccgaagcgga gcataacagg agcggctcat caagaaacat tgcggcgtta
6121 tatcggcatc gacgaacgga gcggaaaaat acagacggtc gtcaaaaaga aactatccga
6181 gatccaactg gataaaacag gtcatttccc aatgtacggg aaagaaagcg atccaaggac
6241 atatgaagcc attcgccaac ggttgcttga acataacaat gacccaaaaa aggcgtttca
6301 agagcctctg tataaaccga agaagaacgg agaactaggt cctatcatcc gaacaatcaa
6361 aatcatcgat acgacaaatc aagttattcc gctcaacgat ggcaaaacag tcgcctacaa
6421 cagcaacatc gtgcgggtcg acgtctttga gaaagatggc aaatattatt gtgtccctat
6481 ctatacaata gatatgatga aagggatctt gccaaacaag gcgatcgagc cgaacaaacc
6541 gtactctgag tggaaggaaa tgacggagga ctatacattc cgattcagtc tatacccaaa
6601 tgatcttatc cgtatcgaat ttccccgaga aaaaacaata aagactgctg tgggggaaga
6661 aatcaaaatt aaggatctgt tcgcctatta tcaaaccatc gactcctcca atggagggtt
6721 aagtttggtt agccatgata acaacttttc gctccgcagc atcggttcaa gaaccctcaa
6781 acgattcgag aaataccaag tagatgtgct aggcaacatc tacaaagtga gaggggaaaa
6841 gagagttggg gtggcgtcat cttctcattc gaaagccggg gaaactatcc gtccgttata
6901 aatcagacaa aatggcctgc ttatgaagcg ggccattttt gtttaatcct gtcgacttgc
6961 cggaattctt tcacagccgc atccctttct tccgcaggca gcgccttcca gcccgcccag
7021 tcgagagaac ggaaatcatg cagtgagtac cagccatcta gtgtttctgc tgcttcgctc
7081 atttttctcc ctccaaatgt ttttctcttc ccttactata acacagttta gaagagggag
7141 aaaatattac ggctgccgcg aaattgtcta aaagaagcct tcttttgcct ttgattttcc
7201 ggaaaaatgg attattctta aaatggatac cgttatacgg atcccatgac gctagtatcc
7261 agctgggtca tagttcccct gagattatcg ctgtggtata atgaaagtta taccacagca
7321 atgatctcag ggttactatg ataagggctt tctgcctata ggcagactga cccgtggcgt
7381 tggggatcgc ctatcgcccg ctttcttcgg gcattcccca ctcttaggcg aacgaagacc
7441 ttggcggaaa acgtcaaggt ctttttt
//
